fer 1 co treatment  (Galectin Therapeutics)

Journal: Cancers

Article Title: Targeting Galectin-1 with Triptolide Induces Ferroptosis in Oral Squamous Cell Carcinoma

doi: 10.3390/cancers18050782

Figure Lengend Snippet: Triptolide (TPL) induces ferroptosis in OSCC cells by promoting lipid peroxidation and suppressing GPX4 expression. ( A ) Chemical structure of Triptolide (TPL). ( B – D ) Cell viability of OSCC cell lines SAS ( B ), SCC25 ( C ), and HSC-3 ( D ) treated with increasing concentrations of TPL (0–80 nM) for 24 and 48 h was measured using the methylene blue assay. ( E ) Lipid peroxidation was assessed using C11-BODIPY 581/591 staining followed by flow cytometry. SAS and HSC-3 cells were treated with TPL (20 nM, 48 h) alone or co-treated with Ferrostatin-1 (Fer-1, 10 μM). Fer-1 was added 1 h prior to TPL treatment. Right panel: quantitative analysis of lipid ROS-positive cells. Letters B denote the flow cytometric gating regions used to identify ROS-positive cell populations. ( F ) Western blot analysis showing GPX4 expression in SAS cells following TPL treatment (0–20 nM, 48 h). GAPDH served as the loading control. Densitometric quantification indicates a dose-dependent reduction in GPX4 expression. The GAPDH blot shown in F is reused in A because both panels were generated from the same experimental membrane under identical conditions. Data represent mean ± SD from three independent biological replicates ( n = 3). * p < 0.05 compared with control. The original western blot figures can be found in .

Article Snippet: Moreover, TPL exposure decreased the expression of both Galectin-1 and GPX4, whereas Fer-1 co-treatment partially restored their levels ( F).

Techniques: Expressing, Staining, Flow Cytometry, Western Blot, Control, Generated, Membrane

Journal: Cancers

Article Title: Targeting Galectin-1 with Triptolide Induces Ferroptosis in Oral Squamous Cell Carcinoma

doi: 10.3390/cancers18050782

Figure Lengend Snippet: Functional involvement of Galectin-1 in TPL-induced ferroptosis in OSCC cells. ( A ) SAS cells were transfected with Galectin-1–targeting siRNA (si-Gal-1) or scrambled control for 48 h, and cell viability was assessed using the methylene blue assay. ( B ) SAS cells were treated with Triptolide (TPL, 20 nM) for 48 h in the presence or absence of Ferrostatin-1 (Fer-1, 10 μM), and cell viability was measured. ( C ) Lipid reactive oxygen species (ROS) levels were determined by C11-BODIPY 581/591 staining followed by flow cytometry in TPL-treated cells (20 nM, 48 h) with or without ectopic Galectin-1 expression achieved by transfection with pCMV-Gal-1. Left panel: representative histograms; right panel: quantitative analysis of lipid ROS–positive cells. Letters B and G denote the flow cytometric gating regions used to identify ROS-positive cell populations. ( D ) Cell viability of TPL-treated cells (20 nM, 48 h) transfected with pCMV-Gal-1 or control vector. ( E ) Western blot analysis of GPX4 expression following Galectin-1 knockdown. ( F ) Western blot analysis of Galectin-1 and GPX4 expression in cells treated with TPL (20 nM, 48 h) alone or in combination with Fer-1 (10 μM). Densitometric analysis was performed using ImageJ software, and protein expression levels were normalized to GAPDH. Data are presented as mean ± SD from three independent biological replicates ( n = 3). * p < 0.05 compared with the respective control groups. The original western blot figures can be found in .

Article Snippet: Moreover, TPL exposure decreased the expression of both Galectin-1 and GPX4, whereas Fer-1 co-treatment partially restored their levels ( F).

Techniques: Functional Assay, Transfection, Control, Staining, Flow Cytometry, Expressing, Plasmid Preparation, Western Blot, Knockdown, Software